Table of Content
List of Contributors XI
About the Series Editors XVII
Preface XIX
Part I Microsystems for Single-Cell Analysis 1
1 Types of Clinical Samples and Cellular Enrichment Strategies 3
Koh Meng Aw Yong, Zeta Tak For Yu, Krystal Huijiao Guan, and Jianping Fu
1.1 Introduction 3
1.2 Types of Clinical Samples 4
1.2.1 Solid Clinical Samples 4
1.2.1.1 Cellular Subtypes Found in Solid Clinical Samples 5
1.2.2 Liquid Clinical Samples and Cellular Subtypes 8
1.2.2.1 Blood 8
1.2.2.2 Bone Marrow 9
1.2.2.3 Placental or Umbilical Cord Blood 10
1.2.2.4 Urine 10
1.2.2.5 Cerebrospinal Fluid (CSF) 10
1.2.2.6 Saliva 11
1.3 Sample Processing and Conventional Methods of Cell Enrichment 11
1.3.1 Processing Solid Clinical Samples 11
1.3.1.1 Processing Liquid Samples 12
1.3.2 Cell Enrichment 12
1.3.2.1 Laser Capture Microdissection (LCM) 12
1.3.2.2 Density Gradient Centrifugation 13
1.3.2.3 Fluorescence-Activated Cell Sorting (FACS) 13
1.3.2.4 Magnetic Activated Cell Sorting (MACS) 15
1.3.2.5 Cell Search TM 15
1.4 Microscale/Nanoscale Devices for Cellular Enrichment 16
1.4.1 Filtration Approaches 16
1.4.2 Hydrodynamic Mechanisms 17
1.4.3 Surface Treatments 19
1.4.4 Magnetophoresis 19
1.4.5 Electrophoresis 20
1.4.6 Acoustophoresis 21
1.4.7 Optical Tweezers/Traps 22
1.5 Conclusion 23
References 23
2 Genome-Wide Analysis of Single Cells and the Role of Microfluidics 29
Sayantan Bose and Peter A. Sims
2.1 Motivation for Single-Cell Analysis of Genomes and Transcriptomes 29
2.2 Single-Cell Genomics 30
2.2.1 Major Technical Challenges 30
2.2.2 Approaches to Single-Cell Genomics 31
2.2.3 The Application and Impact of Microfluidics in Single-Cell Genomics 34
2.3 Single-Cell Transcriptomics 36
2.3.1 Major Technical Challenges 36
2.3.2 Approaches to Single-Cell Transcriptomics 39
2.3.3 Application and Impact of Microfluidics in Single-Cell Transcriptomics 42
2.4 The Future of Genome-Wide Single-Cell Analysis with Microfluidics 45
2.4.1 Recent Advances in the Scalability of Single-Cell Analysis using Microfluidics 45
2.4.2 How Microfluidics will Expand the Application-Space for Single-Cell Analysis 46
2.4.3 Outstanding Hurdles for Genome-Wide Analysis of Single Cells 47
2.4.4 Prospects for Clinical Applications of Microfluidic Single-Cell Analysis 48
Keywords and Definitions 48
References 49
3 Cellular Immunophenotyping: Industrial Technologies and Emerging Tools 57
Kara Brower and Rong Fan
3.1 Cellular Immune Status and Immunophenotyping 57
3.2 Surface Marker Phenotyping 60
3.2.1 Multicolor Flow Cytometry 60
3.2.2 Commercial Flow Cytometers 62
3.2.3 High-Content Imaging Cytometry 63
3.2.4 Current Limitations and Further Development of Flow Cytometry 64
3.3 Functional Phenotyping 65
3.3.1 ELISpot Technologies 66
3.3.2 Multiplexed Immunoassays 67
3.3.3 Emerging Single-Cell Technologies 68
3.4 Conclusion 70
Keywords and Definitions 71
References 71
4 Microsystem Assays for Studying the Interactions between Single Cells 75
Vandana Kaul and Navin Varadarajan
4.1 Introduction 75
4.2 Advantages of Single-Cell Analysis over Conventional Assay Systems 80
4.3 Analysis of Cell–Cell Communication between Pairs of Single Cells 81
4.3.1 Integrated Microfluidic Coculture Systems and Microwell Arrays 81
4.3.1.1 Microengraving 81
4.3.1.2 T-Cell Proliferation 82
4.3.1.3 T-Cell Cytotoxicity 82
4.3.1.4 NK-Cell Cytotoxicity 84
4.3.1.5 High-Throughput Stem Cell Coculture Array 84
4.3.1.6 Microfluidics-Based Single-Cell RNA-seq for Intercellular Communication 85
4.3.1.7 Single-Cell Signaling Chip 85
4.3.2 DEP Arrays 87
4.3.2.1 Tumor Cell–Endothelial Cell Interaction 87
4.3.2.2 Immune-Cell Cytotoxicity 88
4.3.3 Microfluidic Hydrodynamic Trapping 89
4.3.3.1 Sequential Hydrodynamic Trapping Device 89
4.3.3.2 Intercellular Communication via Gap Junctions 89
4.3.3.3 Cell–Cell Fusion 90
4.3.4 Optical Methods 91
4.3.4.1 Laser-Guided Cell Micropatterning 91
4.3.4.2 Optical Tweezers 91
4.3.4.3 Optoelectronic Tweezers 93
4.3.5 Magnetic Methods 93
4.3.5.1 Magnetic Pattern Arrays 94
4.3.5.2 Magnetic Microflaps 94
4.3.6 Acoustic Methods 94
4.3.6.1 Ultrasonic Standing Waves (USWs) for 2D and 3D Cell–Cell Interaction 95
4.3.6.2 Standing Surface Acoustic Waves for Cell Patterning 96
4.3.6.3 Ultrasonic-Based Method for Cell–Cell Interactions in Microwell Arrays 96
4.4 Conclusions 97
Acknowledgments 98
References 98
5 Modeling Microvascular Disease 105
Hope K.A. Gole and Wilbur A. Lam
5.1 Introduction 105
5.2 Microvascular Disease 106
5.3 Macromodeling 107
5.4 Micromodeling 109
5.4.1 Fabrication 110
5.4.2 Design and General Applications 112
5.4.3 Disease-Specific Applications 115
5.4.4 Advantages and Disadvantages 120
5.5 Summary 122
References 122
Part II Tiny Technologies for Modulating Biological Systems 127
6 Nanotechnologies for the Bioelectronic Interface 129
Benjamin W. Avants, Hongkun Park, and Jacob T. Robinson
6.1 Introduction 129
6.2 Modeling the Bioelectronic Interface 130
6.3 Experimental Approaches for Extra-Cellular Coupling 132
6.4 State-of-the-Art Extra-Cellular Nanoscale Interfaces 133
6.5 Experimental Approaches for Intra-Cellular Coupling 134
6.6 State-of-the-Art Intra-Cellular Nanoscale Interfaces 135
6.7 Experimental Approaches for In-Cell Coupling 137
6.8 Outlook 138
References 139
7 Intracellular Delivery of Biomolecules by Mechanical Deformation 143
Armon Sharei, Shirley Mao, Robert Langer, and Klavs F. Jensen
7.1 Introduction 143
7.2 Delivery Concept 148
7.2.1 Design 149
7.2.2 Governing Parameters 150
7.3 Cytosolic Delivery by Diffusion 151
7.3.1 Modeling Diffusion 153
7.3.2 Imaging of Membrane Disruptions 157
7.4 Applicability across Cell Types and Delivery Materials 158
7.4.1 Flexibility in Addressing Different Delivery Material 162
7.4.2 Enabling New Research and Clinical Applications 164
7.4.2.1 Cell Reprogramming 164
7.4.2.2 Quantum Dot delivery 166
7.4.2.3 Immune Cell Delivery 166
7.5 Summary 167
7.6 Appendix 169
7.6.1 Device Design Guidelines for New Cell Types 169
7.6.2 Design Parameters 169
7.6.3 Device Nomenclature 170
7.6.4 Defining Delivery Efficiency 171
7.6.5 Device Recovery 171
7.6.6 Reagent Use 171
Acknowledgments 173
Conflict of Interest 173
Keywords and Definitions 174
References 174
8 Microfluidics for Studying Pharmacodynamics of Antibiotics 177
Ritika Mohan, Amit V. Desai, Chotitath Sanpitakseree, and Paul J.A. Kenis
8.1 Background on Antibiotic Resistance 177
8.2 Methods for Antibiotic Susceptibility Testing (AST) 178
8.2.1 Conventional Methods 178
8.2.2 Integrated Microfluidic-Based Approaches 179
8.2.3 Translation of Microfluidic-Based Approaches 182
8.3 Applying Pharmacokinetics/Pharmacodynamics to AST 184
8.3.1 Significance of PK/PD 184
8.3.2 Advantages of Microfluidic-Based Approaches for PK/PD Analysis 185
8.4 Application of Microfluidic-Based Approach for PK/PD Modeling 185
8.4.1 PD Modeling 186
8.4.1.1 Monomicrobial Cultures: MIC Determination of E. coli against Amikacin 188
8.4.1.2 Polymicrobial AST: MIC Determination of E. coli and P. aeruginosa against Amikacin 189
8.4.2 PK Modeling 192
8.5 Summary and Future Outlook 194
Acknowledgments 196
References 196
9 Microsystems Models of Pathophysiology 203
Marie-Elena Brett and David K.Wood
9.1 Vascular and Hematologic Pathologies 205
9.1.1 Thrombosis 205
9.1.2 Sickle Cell Disease 208
9.1.3 Malaria 212
9.1.4 Atherosclerosis 213
9.1.5 Model Limitations and Future Opportunities 214
9.2 Organ-Specific Pathologies 217
9.2.1 Lung 218
9.2.2 Brain 220
9.2.3 Kidney 222
9.2.4 Liver 224
9.2.5 Challenges and Opportunities 226
9.2.5.1 Considerations and Challenges 227
9.2.5.2 Opportunities 230
9.3 Cancer 230
9.3.1 Microscale Tumor Models 231
9.3.2 Metastasis 232
9.3.3 Drug Delivery and Pharmacokinetics 236
9.4 Summary 237
References 238
10 Microfluidic Systems for Whole-Animal Screening with C. elegans 245
Navid Ghorashian, Sertan Kutal Gökçe, and Adela Ben-Yakar
10.1 Importance 245
10.2 Introduction 245
10.3 A Versatile Animal Model: Caenorhabditis elegans (C. elegans) 246
10.3.1 C. elegans Culturing Techniques 247
10.3.2 C. elegans as a Model of Neurological Disease 247
10.3.3 C. elegans as a Drug-Screening Model 249
10.3.4 Current State of the Art in Automated C. elegans Screening 249
10.4 Microfluidics 251
10.4.1 Microfluidic Device Fabrication 251
10.4.2 Fluid Dynamics Modeling in Microfluidics 252
10.4.3 Microfluidics Interfacing with Multiwell Plates 255
10.4.4 Microfluidic Flow Control and Valve Multiplexing 255
10.5 Microfluidics for C. elegans Biology 257
10.5.1 Microfluidic Worm Immobilization and High-Resolution Optical Interrogation Platforms 257
10.5.1.1 Single Trap Microfluidic Platforms for Worm Processing One at a Time 258
10.5.1.2 Multitrap Microfluidic Platforms to Enable Parallel Worm Processing 262
10.5.2 Microfluidic Population Delivery for Serial Processing 264
10.6 Conclusions and Future Directions 266
Author Contributions 266
References 266
Index 273
